Elimination and reconstitution of the requirement for hormone in promoting temperature-dependent transformation of cytosolic glucocorticoid receptors to the DNA-binding state

Cytosols contain a heat-stable, chelatable, anionic, molybdate-like factor that stabilizes glucocorticoid receptors in a heteromeric complex with hsp90 (refers to the 90-kDa heat shock protein) and inhibits their transformation to the DNA-binding state (Meshinchi, S., Grippo, J.F., Sanchez, E.R., Bresnick, E.H., and Pratt, W.B. (1988) J. Biol. Chem. 263, 16809-16817). In this work, we demonstrate that removal of this factor by passage of L cell cytosol through the metal-chelating resin Chelex-100 makes the glucocorticoid receptor unstable, thus markedly facilitating both its dissociation from hsp90 and its transformation to the DNA-binding state. In normal cytosol, both temperature-mediated dissociation of hsp90 and temperature-mediated receptor transformation are hormone-dependent events. In the Chelex-treated, metal-depleted cytosol, however, temperature-mediated dissociation of hsp90 and receptor transformation occur very rapidly in a manner that is no longer hormone-dependent. When boiled L cell cytosol is added to the metal-depleted receptor system, the hormone dependence of both temperature-mediated dissociation of receptor from hsp90 and receptor transformation to the DNA-binding state is reconstituted. Like boiled cytosol, molybdate stabilizes the receptor complex and inhibits its transformation in metal-depleted cytosol, but it does not reconstitute the hormone dependence of the system. These results support the proposal that an endogenous metal anion interacts with the glucocorticoid receptor to stabilize it in the heteromeric, inactive, non-DNA-binding state in cytosol and that binding of the hormone promotes conversion of the receptor to the DNA-binding state through an effect on this metal anion center.     

In vitro meristem culture of M.4 apple (Malus pumila Mill.). I. Optimal nutrient medium

Shoot tips of M.4 apple clone were excised from actively growing one year-old stoolbed branches, and cultured in order to determine the optimal nutrient medium for each stage of their in vitro culture. The basal medium (BM) used was that described by Murashige and Skoog, supplemented with vitamins, glycine, myoinositol, sucrose, with or without agar, and different combinations of plant growth regulators. Best media for each stage were: BM+0.5 mg 1^-1 indole-3yl-butyric acid (IBA)+0.5 mg 1^-1 6-benzylaminopurine (BAP) for explant establishment (Stage I); BM+0.1 mg 1^-1 IBA+1.0 mg 1^-1 BAP for multiplication and internode enlargement (Stage II); and 2.0 mg 1^-1 IBA+0.1 mg 1^-1 BAP without agar for the rooting of the plantlets (Stage III).     

Incorporation of free fatty acids into acylthioesters and lipids of developing sunflower seeds

The synthesis of lipids from radioactive fatty acids in developing sunflower seeds has been examined. Lauric, palmitic, stearic and oleic acids were us     

Hypertrophic cardiomyopathy associated with left ventricular aneurysm and normal coronary arteries: Case study indicating genetic tendencies of cardiomyopathy

A 50-year-old man presented with hypertrophic obstructive cardiomyopathy (HOC) associated with a left ventricular aneurysm and normal coronary arteries. His history revealed no evidence of myocardial infarction or atypical angina. Physical examination disclosed HOC but did not suggest the presence of an aneurysm. Although the patient was treated medically, heart failure ensued, and he died suddenly while working his farm. Subsequent investigation of the patient's family revealed that three of his five children were also affected by cardiomyopathy, which was especially pronounced in the eldest, a 22-year-old man. The possible hemodynamic relationship between HOC and left ventricular aneurysm is discussed, along with probable indications. The role of left ventricular aneurysm is also presented in relation to the natural history of the disease.     

Selective enrichment of aromatic amino acid auxotrophs in Hansenula polymorpha.

Aromatic amino acid auxotrophs of the methanol-utilizing yeast Hansenula polymorpha were effectively selected by the use of nystatin and a medium that inhibits the growth of tyrosine auxotrophs. The procedure resulted in a frequency of aromatic auxotrophs of 2% of survivors and an enrichment of 20-fold. The new procedure also takes less time than traditional procedures. Of the auxotrophic mutants isolated, two-thirds required tyrosine and the remainder were tyrosine-phenylalanine double auxotrophs.     

Simple agarose gel electrophoretic method for the identification and characterization of plasmid deoxyribonucleic acid.

Agarose gel electrophoresis may be employed effectively for the detection and preliminary characterization of plasmid deoxyribonucleic acid (DNA) present in clinical isolates and laboratory strains of gram-negative microorganisms. The method is sensitive and does not require radioisotopes or ultracentrifugation. The estimation of plasmid mass from the extent of DNA migration in gels compares favorably with results obtained by electron microscopy of plasmid DNA purified by equilibrium density centrifugation. The method has proved to be a useful tool for survey work and the epidemiological investigation of plasmid dissemination, as well as an important adjunct to the genetic analysis of plasmids.     

Recurrences and Human Agential Meaning Grounding: Laying a Path in Walking

This article addresses the semiotic problem of how meaning is agentially grounded: how actual meaning is possible and is justifiably supported by agents’ capabilities and purposes. This article is particularly focused on human agential grounding; however, to a great degree, insights presented here can be extended to other living beings. Specifically, agential meaning is examined here inside the framework of agentive semiotics and embodied, situated and enactive cognition theories, in line with the mind-life continuity general thesis (which intends to naturalize mind and experience). To offer clarity and methodological precision about agential grounding, three explanation categories (called recurrences) are proposed: phylogenetic recurrence, the evolutionary basis for corporal/embodied grounding; ontogenetic recurrence, the developmental basis for individual meaning grounding; and collective recurrence, the basis for meaning recognized, attributed and assigned inside social contexts. These recurrences are conceived as three types of general processes that constantly enclose possibilities for purpose and meaning emergence in humans. As a result of these types of recurrences, two categories of human agendas or purposes are also proposed: individual and collective. Finally, remarks about how these categories can be useful for semiotic analysis and further research are suggested.     

A Lead ANRIL Polymorphism Is Associated with Elevated CRP Levels in Periodontitis: A Pilot Case-Control Study

Elevated high sensitive C-reactive protein (hsCRP) is a marker for systemic inflammation and a risk marker for atherosclerotic cardiovascular disease (ACVD), and has also been associated with periodontitis. Inter-individual variation for hsCRP in periodontitis has been shown. ANRIL is the strongest genetic susceptibility locus for both periodontitis and ACVD, and it is speculated that genetic variation in ANRIL may modulate inflammatory processes. Therefore, we explored the possible association between hsCRP plasma levels and a leading ANRIL single nucleotide polymorphism (SNP) in periodontitis patients and controls. 171 healthy subjects with North European descent (115 periodontitis and 56 controls) were included in this case-control study. hsCRP levels were determined and subjects were genotyped for the leading ANRIL SNP rs1333048. In a multivariate analysis, periodontitis, female gender, increasing BMI and homozygosity for the major allele (AA-genotype) of rs1333048 were significantly associated with elevated hsCRP plasma levels (p = 0.012, p = 0.004, p = 0.007 and p = 0.003, respectively). Periodontitis patients with rs1333048 AA-genotype showed higher levels of hsCRP than those carrying the minor C allele (median: 4.5 mg/L vs. 1.6 mg/L, p_adjusted = 0.007). This study is the first to show that, in addition to gender and BMI, also a leading SNP in ANRIL is explanatory for inter-individual variation in hsCRP levels in periodontitis patients of North European descent.     

Effect of the hydraulic retention time on the anaerobic biofilm reactor efficiency applied to screened cattle waste treatment

Summary It was found that the increase of the hydraulic retention time improved the organic matter removal and the conversion of organic to ammonia nitrogen. Methane concentration in the biogas was not affected by the hydraulic retention time.The reactor in operation during one year was not clogged.